Introduction

Nonalcoholic fatty liver disease (NAFLD) is the most common cause of chronic liver disease worldwide, with a prevalence of 25% in the adult population. NAFLD encompasses a wide spectrum of histological conditions, from simple steatosis to nonalcoholic steatohepatitis (NASH), fibrosis and cirrhosis. Inflammation is one of the principal hallmarks of NAFLD. Polyclonal free light chains (FLCs) λ and κ reflect B-cell activation and high serum levels of FLCs are observed in a variety of inflammatory conditions.

Aims & Methods

The aim of this study is to evaluate the potential role of FLCs as biomarkers of inflammation and fibrosis in NAFLD/NASH patients. 254 patients with metabolic liver disease were consecutively enrolled in the study. 89 patients had simple steatosis, 88 had NASH, 77 had liver cirrhosis (45 with clinically significant portal hypertension [CSPH]). Diagnosis of NASH was histologically assessed. 101 patients had compensated advanced chronic liver disease (cACLD) based on histology (F>3) or elastography (liver stiffness measurement>15 kPa). Medical and pharmacological anamnesis, anthropometric measurements and laboratory tests (included λ and κ FLCs) were obtained for all patients.

Results

The concentration of total FLCs levels in patients with cirrhosis were higher compared with patients with NAFLD/NASH (p<0.01), while no significant difference in λ and κ FLCs levels were observed between NASH and NAFLD patients. Among patients who underwent biopsy, λFLCs tended to increase gradually with lobular inflammation severity, and patients with advanced fibrosis (F3-4) had a significantly higher FLCs compared with patients with mild-to-moderate fibrosis (F<3; p<0.05). Patients with clinically significant portal hypertension (CSPH) showed higher levels of FLCs compared to patients without CSPH (p<0.01). At multivariate logistic analysis both kFLCs and λFLCs are significantly associated with diagnosis of CSPH and cACLD, independently from age, sex, diabetes, dyslipidemia and obesity. Diagnostic performance of cACLD and CSPH were high for both kFLCs (AUC 0.79 and 0.87 respectively) and λFLCs (AUC 0.81 and 0.89 respectively).

Conclusion

This study highlights the role of adaptive immune system in the pathogenesis of NAFLD. FLCs may be considered useful biomarkers of advanced liver disease (cACLD and CSPH) in patients with NAFLD.

Introduction

The part of NAFLD children has a progressive sub-phenotype known as nonalcoholic steatohepatitis (NASH), characterized by the ability to develop advanced fibrosis, cirrhosis, and end-stage liver disease [1]. Given the high prevalence of NAFLD and invasiveness of the liver biopsy, there is an essential need to find safe non-invasive modalities to determine liver fibrosis in children [2].

Aims & Methods

To investigate serum fibrosis markers in obese children with non-alcoholic fatty liver disease (NAFLD) depending on the presence of liver fibrosis.
94 children aged 9 to 17 years (average age 12.15 ± 2.51 years) were included in the study. According to the transient elastography data (FibroScan®502touch, Echosence, France) and body mass index, children were divided into four groups: group 1 - 27 obese NAFLD children with fibrosis ≥F1, group 2 - 35 obese NAFLD children without fibrosis, group 3 - 18 obese children without NAFLD and fibrosis. Group 4 (control) consisted of 14 children with normal weight without NAFLD and liver fibrosis. The serum levels of vascular endothelial growth factor (VEGF) (Wuhan Fine Biotech Co Ltd, China), transforming growth factor-beta 1 (TGF-β1) (IBL International, Germany), cytokeratin-18 (CK-18) (IDL Biotech AB, Sweden) were determined by enzyme-linked immunosorbent assay (ELISA). Serum content of hydroxyproline free (HPf) and hydroxyproline protein-bound (HPp/b) according to Osadchuk and glycosaminoglycans (GAG) according to Rimington were evaluated.

Results

In 1 group children, a significant increase in the median level of CK-18 (in 2.2 times, p<0.05), VEGF (in 2.4 times, p<0.05), and TGF-β1 (in 2.7 times, p<0.05) was found compared to 4 group children. Moreover, CK-18 level was significantly higher in 1 group of patients compared to 2 group (in 1.4 times, p<0.05) and 3 group (in 1.8 times, p<0.05). In all group children, an increase in the serum HPp/b content was found: in 1 group - in 1.2 times (p<0.01), in 2 group - in 1.2 times (p<0.05), in 3 group - in 1.3 times (p<0.05) compared to 4 group. The changes in the content of hydroxyproline led to an increase in the HPp/b/HPf ratio in all groups: 1.4 times (p<0.01) in 1 and 2 groups, in 1.3 times (p<0.01) in 3 group. Also, in 1 group of children median GAG level was increased by 1.3 times (p<0.001).
In children with NAFLD HPf content negatively correlated with CAP (r= - 0.408; p<0.01), CK-18 (r=-0.469; p<0.01), the ratio HPp/b/HPf positively correlated with CAP (r=0.307; p<0.05). CK-18 positively correlated with fibrosis degree (according to METAVIR) (r=0.468; p=0.008) and liver steatosis degree (r=0.357; p=0.048). A positive correlation was also found between VEGF levels and the degree of liver fibrosis (r=0.372, p=0.036).

Conclusion

Serum levels of VEGF, TGF-β1, HPp/b, and GAG were significantly elevated in children with non-alcoholic fatty liver fibrosis. Assessment of these parameters may be useful for early diagnosis of liver fibrosis in children with NAFLD.

References

1. Schwimmer JB, Johnson JS, Angeles JE, et al. Microbiome Signatures Associated With Steatohepatitis and Moderate to Severe Fibrosis in Children With Nonalcoholic Fatty Liver Disease. Gastroenterology. 2019;157(4):1109-1122. doi:10.1053/j.gastro.2019.06.028.
2. Alkhouri N. Putting it all together: Noninvasive diagnosis of fibrosis in nonalcoholic fatty liver disease in adults and children. Clin Liver Dis (Hoboken). 2017;9(6):134-137. Published 2017 Jun 29. doi:10.1002/cld.636

Introduction

Non-alcoholic fatty liver disease (NAFLD) can progress to fibrosis and cirrhosis. The evaluation of fibrosis in chronic liver disease using non-invasive methods is a current topic. Standard methods are liver stiffness measurement and serum biomarkers. Splenic elasticity measurement is a recent and interesting non-invasive approach that arouses the interest of the latest studies to evaluate fibrosis severity in NAFLD.
The aim of this study is to identify patients with NAFLD and advanced liver fibrosis, by establishing correlations between splenic and liver elasticity and fibrosis serum biomarkers.

Aims & Methods

This is a prospective study conducted in the hepato-gastroenterology department including all patients followed for hepatic steatosis during the period from July 2022 to January 2023. Patients with a family or personal history of chronic liver disease, and women with acute steatohepatitis in pregnancy or cholestasis in pregnancy were excluded. Measurement of hepatic and splenic stiffness and the degree of hepatic steatosis was done using liver elastrography (Fibroscan®) and Transient elastography with controlled attenuation parameter (CAP), respectively. NAFLD fibrosis (NFS) and Fibrosis-4 scores were calculated. Data were entered and processed using SPSS software version 22.0.

Results

We included 44 patients, divided into 28 women and 16 men (Sex-ratio F/M = 1.75), with a mean age of 61.4 years [32 to 84 years]. Three patients were smokers and four were alcoholics. Hypertension was found in 56.8%, 54.5% were diabetics and 52.3% were dyslipidemics. The mean BMI was 29.6 kg/m²±4.15 kg/m². Obesity was found in 45.5%, 36.4% were overweight and 18.2% of patients were normal weight. Hepatic steatosis was discovered following pain in the right hypochondrium in 79.5% of the cases and in front of a cytolysis in the remaining cases. All patients had diffuse steatosis without portal hypertension or degeneration signs on ultrasound.
Biological results showed cytolysis in seven patients. No patient had cholestasis. All patients had preserved liver function: normal prothrombin level and albumin. Hyperglycemia and lipid disturbance were observed in 43.2% and 36.4% of cases respectively.
Fibrosis was assessed by FIB4 score with a mean value of 1.11± 0.78; NFS score with a mean value of -1.16±1.18 and liver stiffness ranging from 2.4Kpa to 28.4Kpa (mean value 6.67Kpa). The degree of fibrosis judged by Fibroscan results was F0-F1 in 31 cases, F2 in 5 cases, F3 in 5 cases and F4 in 3 cases. Advanced fibrosis was correlated with NFS score (p=0.016), FIB-4 (p=0.001); but not associated with CAP value (p=0.51).
The mean splenic elasticity was 27.57 Kpa (ranging from 8.3 to 75 Kpa). It was significantly associated with liver elasticity (p=0.012) with an AUROC of 0.784. A threshold of 17.4 Kpa was predictive of advanced fibrosis with a specificity of 85.7% and a sensitivity of 73.5% (CI [0.029-0.404]). Similarly, splenic elasticity was correlated with CAP value (p=0.013). No association was identified with NFS and FIB4 scores.

Conclusion

Measurement of splenic elasticity is a simple and reliable non-invasive tool, comparable to liver elasticity, for the prediction of liver fibrosis during NAFLD.

Disclosure

no conflict of interest

Introduction

Currently, about 25-30% of the world population is affected by non-alcoholic fatty liver disease (NAFLD); over time, about 20% tends to develop a certain degree of inflammation in the liver tissue, defying the non-alcoholic steatohepatitis (NASH). Moreover, approximately 20% of NASH patients evolve into a high-grade liver fibrosis condition that leads to severe liver impairment (cirrhosis) (1, 2, 3).
The serial use of non-invasive fibrosis tests can refine prognosis in patients with non-alcoholic fatty liver disease (NAFLD) and evaluate the progression or improvement of liver fibrosis.

Aims & Methods

We evaluated predictors of improvement or worsening of liver stiffness measurements (LSM) in well characterized cohorts of patients with NAFLD from London (UK) and Milan (Italy).
We included two consecutive cohorts of 405 patients with at least two outpatient visits between 2014 and 2022. The minimum time interval between baseline and follow-up LSM was >6 months. LSM worsening was defined as an increase of >20% kPa if the baseline LSM was ≥5 kPa, or a follow-up LSM >6 kPa if the baseline LSM was <5 kPa. An LSM improvement was defined as an LSM decrease of >20% kPa (if the baseline LSM was >6 kPa).
A significant change in weight was defined as a >5% reduction or increase at follow-up, while a significant change in glycated hemoglobin (HbA1c) was defined as a >10% reduction or increase.
The variation between the true and expected FIB-4 index (based on the patient’s age at the follow-up visit, but using the blood tests performed at the first visit) was calculated. A significant improvement or worsening in FIB-4 was defined as a >20% variation between the actual and “expected” FIB-4.

Results

Of the 405 patients, 282 (70%) were males; mean age was 54±11 years. The median time from the first visits was 20.3 (13.9 – 28.7) months.
128 and 54 patients had an LSM >8 kPa and LSM>12 KPa at follow-up, respectively.
77 patients (19.3%) had an LSM improvement, while 67 (16.8%) had an LSM worsening; 256 patients (63.9%) maintained a stable value.
In patients with an LSM improvement, 22 had an improvement, 12 had worsening, while 37 had stable FIB-4.
In patients with an LSM worsening, 10 had an improvement, 20 had worsening, while 33 had a stable FIB-4.
In multiple logistic regression analysis, LSM worsening was independently associated with an increase in ALT and in HbA1c levels (OR 1.02 and 1.03 respectively), but also with a reduction in total cholesterol and AST levels (OR 0.98 for both). LSM improvement was independently associated with weight and ALT reduction (OR 0.93 and 0.98 respectively), but also with an increased AST level (OR 1.02).

Conclusion

More than 35% of patients with NAFLD have significant changes in their LSM measurements over a period of 20 months, with worsening or improvement at similar rates. Improvement in weight is independently associated with significant improvement in LSM measurements. On the converse, worsening in metabolic comorbidities, particularly glycemic control, could be associated with significant worsening in LSM, further supporting a multidisciplinary model of care.

References

1. Younossi Z, et al. Global burden of NAFLD and NASH: trends, predictions, risk factors and prevention. Nat Rev Gastroenterol Hepatol 2018;15(1):11-20.
2. Sheka AC, et al. Nonalcoholic Steatohepatitis: A Review. JAMA 2020;323(12):1175-1183.
3. Liu J, et al. Estimating global prevalence, incidence, and outcomes of non-alcoholic fatty liver disease from 2000 to 2021: systematic review and meta-analysis. Chin Med J (Engl). 2022 Jul 20;135(14):1682-1691.

Introduction

As the most common form of chronic liver disease, the increasing prevalence of non-alcoholic fatty liver disease (NAFLD) is a global concern. NAFLD and obesity are independently related to an increased risk for atherosclerotic cardiovascular disease (ASCVD), the primary cause of mortality in NAFLD patients. Even though many subjects with NAFLD are normal weight, it still remains uncertain whether their ASCVD risk is of major importance.

Aims & Methods

The aim of this study is to assess and compare the ASCVD risk between lean and obese patients with NAFLD. Normal weight and obese patients were evaluated between January 2020 and February 2023 and their data was analyzed. NAFLD was diagnosed by vibration-controlled transient elastography (VCTE) with controlled attenuation parameter (CAP) and the American College of Cardiology/American Heart Association guidelines was used to evaluate the ASCVD risk. Moreover, medical history and biochemical data were collected.

Results

In total, 221 patients were included in the final analysis. In the obese group, 164 (63.8%) patients were diagnosed with NAFLD, compared to 57 (12.3%) in the lean population. In comparison to those with obese NAFLD, subjects with lean NAFLD had significantly higher ASCVD scores (mean 15.3% vs 22.7%, p<0.001). Moreover, subjects with lean NAFLD and significant liver fibrosis had a higher risk of ASCVD events, compared to their obese counterparts (OR, 2.51 vs 1.95, p=0.034). On the other hand, in terms of liver stiffness measurements, the prevalence of at least significant liver fibrosis was substantially higher in the obese group. Regarding the presence of the components of the metabolic syndrome, the prevalence of type 2 diabetes mellitus was higher the obese group, while changes in the lipid profile and higher systolic blood pressure were more frequently found in lean subjects.

Conclusion

The ASCVD scores of subjects with lean NAFLD were significantly higher than those with obese NAFLD. The presence of significant liver fibrosis was a notable risk factor associated with an ASCVD event in both study groups, but the effects were more pronounced in individuals with a low body mass index.​